聚甲基丙烯酸酯纳米粒作为反义寡核苷酸传递系统的研究聚甲基丙烯酸酯纳米粒作为反义寡核苷酸传递系统的研究

目的 研究聚甲基丙烯酸酯阳离子纳米粒作为反义寡核苷酸传递系统的可行性。方法 以Eudragit RL100和RS100为材料,采用溶剂-非溶剂法制备纳米粒,再与寡核苷酸混合即得载药纳米粒。用透射电镜观察其形态、粒径测定仪测定粒径、超滤法测定药物的包封率,通过台盼蓝拒染试验和红细胞溶血试验测定纳米粒的细胞毒性,用流式细胞仪测定荧光标记的寡核苷酸的入胞量。结果 纳米粒的形态规整、大小均匀,平均粒径为127 nm左右,几乎所有的药物被负载。使用聚甲基丙烯酸酯纳米粒作为反义寡核苷酸载体后,进入细胞内的药物量急剧增加,并有显著的剂量依赖性,但对细胞有轻微的毒性作用。结论 聚甲基丙烯酸酯阳离子纳米粒是一种稳定、高效的反义寡核苷酸传递系统。

Study on polymethacrylate nanoparticles as delivery system of antisense oligodeoxynucleotides

AIM: To investigate the possibility of polymethacrylate nanoparticles (NP) for antisense oligodeoxynucleotides delivery system. METHODS: The nanoparticles were prepared by evaporating ethenol solution containing Eudragit RL100 or RS100, and then mixtured with oligonucleotides. The morphology and size were investigated by a transmission electron microscope and Mastersizer particle characterization systems, and the cytotoxicity was evaluated by Trypan Blue staining and hemolysis test. The flow cytometer was used to determine the uptake of fluorescence-labelled oligodeoxynucleotides. RESULTS: The morphology of nanoparticles showed spherical and orderly, the average diameter was about 127 nm, and almost the antisense oligodeoxynucleotides (ODN) were loaded when NP: ODN was 6.6. The uptake of ODN was significantly increased when loaded by nanoparticles, which well depended on the nanoparticles concentration. Meanwhile, slightly cytotoxicity was observed when high dose of nanoparticles was used. CONCLUSION: The polymethacrylate nanoparticles appeared to be a promising vehicle for gene delivery.